1. Technology schematic for phenotypic tracking of single molecularly defined B-lineage clones.
Technology schematic for phenotypic tracking of single molecularly defined B-lineage clones. A, Single B-lineage cells identified by CD19 and CD38 characteristics were index sorted into 96-well plates using a 13-parameter FACS panel (see Supplementary Fig. S1 for detailed gating strategy). Single-cell light chain mRNA was reverse transcribed, amplified, barcoded, and sequenced. In parallel, FACS data were visualized as t-SNE maps and single monoclonal B-lineage cells of the predominant clone were mapped onto the t-SNE plots (black) to visualize their phenotypic distribution. B, Illustration of the amplification and barcoding strategy. For each single cell, a first amplification is followed by a nested second amplification that introduces plate-, column-, and well-specific barcodes and the first part of the Illumina adapter sequences. The remaining parts of the adapter sequences were attached in a third PCR reaction (not shown in the figure; see Supplementary Table S4 for primer sequences).
Leo Hansmann et al. Cancer Immunol Res 2017;5:
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